title: Isolation of side population cells from endometrial cancer cells using a violet laser diode.
abstract: Cancer stem cells (CSCs) possess the ability for self-renewal, differentiation, and tumorigenesis and play a role in cancer recurrence and metastasis. CSCs are usually sorted in analysis into side population (SP) cells using ultraviolet (UV) laser (350 nm) excitation; they cannot be stained with Hoechst 33342 because of their efflux ability. However, it is difficult to avoid cell damage using a UV laser. Therefore, we attempted to isolate CSCs using a violet laser (407 nm) excitation to avoid cellular DNA damage. We sorted SP cells and main population (MP) cells from a human endometrial cancer cell line using the FACSAria system equipped with a violet laser and analyzed the biological properties of these cells. SP cells exhibited drug efflux, self-renewal, differentiation abilities, and tumorigenicity. It was found that v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) expression was significantly higher in SP cells than in MP cells. Our results suggest that CSCs exist in the SP fraction sorted using the FACSAria system equipped with a violet laser, which presents a useful tool to isolate small populations of viable putative CSCs from solid tumors and can be used to identify and characterize CSCs.
authors.affiliation: Department of Chemical Technology, Graduate School of Science and Industrial Technology, Kurashiki University of Science and the Arts, Kurashiki, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tomiyasu
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Miyamoto
authors.forename: Michihiro
authors.initials: M
authors.lastname: Mori
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yaguchi
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Yakushiji
authors.forename: Setsuyo
authors.initials: S
authors.lastname: Ohno
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Miyake
authors.forename: Takuya
authors.initials: T
authors.lastname: Sakaguchi
authors.forename: Masatsugu
authors.initials: M
authors.lastname: Ueda
authors.forename: Eiji
authors.initials: E
authors.lastname: Ohno
mesh_terms: D000818:Animals; D018269:Carcinoma, Endometrioid / Q000235:genetics* / Q000473:pathology*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002471:Cell Transformation, Neoplastic; D016889:Endometrial Neoplasms / Q000235:genetics / Q000473:pathology*; D005260:Female; D015870:Gene Expression; D006801:Humans; D054023:Lasers, Semiconductor; D009154:Mutation; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000473:pathology; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D051381:Rats; D058985:Side-Population Cells / Q000166:cytology / Q000473:pathology; D014466:Ultraviolet Rays
chemical_list: C528430:Kras protein, rat; D016283:Proto-Oncogene Proteins p21(ras)